ABSTRACT
OBJECTIVE:To explore basic technology for synthesis of active ingredients of Ophiocordyceps xuefengensis,and provide necessary technical support for comprehensive development of O. xuefengensis sourse. METHODS:Submerged fermenta-tion method was used to cultivate the mycelium,achieving efficient synthesis of active ingredients by controlling medium composi-tion and cultivation conditions. Using the bacteria as starting strain,the effects of different carbon sources (sucrose,glucose and soluble starch),different nitrogen sources (peptone,yeast extract powder,yeast extract,sodium nitrate,potassium nitrate and urea),different vitamin B(vitamin B1 and vitamin B complex)and different initial pH(pH was set at 4,5,6,7,8 and 9,re-spectively)on mycelial growth,extracellular and intracellular polysaccharide synthesis,cordycepin synthesis and intracellular triter-penoid synthesis were investigated to screen the optimal medium composition. RESULTS:The optimal carbon source,nitrogen source,vitamin B and initial pH were sucrose,yeast extract powder,vitamin B1 and 8,respectively. High biomass and metabolite accumulation levels can be obtained when carbon source was sucrose,nitrogen source was yeast extract powder,adding 0.1 g/L vi-tamin B1 with initial pH of 8. CONCLUSIONS:O. xuefengensis can efficiently accumulate metabolites,and achieve the optimiza-tion of strain cell growth and synthesis of active metabolite by optimizing and controlling the fermentation process.
ABSTRACT
Objective:To study the effects of Ophiocordyceps xuefengensis on proliferation of DC -CIK cells and the activity of killing HepG-2 cells by DC-CIK cells.Methods:Peripheral blood mononuclear cells were routinely isolated and induced into DC and CIK.DC and CIK co-cultured by 1∶5 for 7 days,then Ophiocordyceps xuefengensis were added into medicine group ,observed the mor-phology of the cells on the tenth day and counted the DC-CIK number of each group.DC-CIK cells acted as effector cells and the HepG-2 cells as target cells , cck-8 method for the detection of DC-CIK in the killing rate of HepG-2.Results: The Ophiocordyceps xuefengensis was able to proliferate the DC-CIK dramatically ,the optimal concentration was 0.1 mg/ml.Cultivation of Ophiocordyceps xuefengensis induced DC-CIK cells on HepG-2 cells killing effect was better than that of the routine method of DC-CIK cells; the effection of Ophiocordyceps xuefengensis killing HepG-2 cells was not obviously.Conclusion: Ophiocordyceps xuefengensis can enhance the anti-tumor activity of DC-CIK mainly by promoting the proliferation of it.
ABSTRACT
Using the HPLC-Q-TOF-MS/MS to analyze and identify the main components in the ethanol extract of Ophiocordyceps xuefengensis sp. nov. The ethanol extract of O. xuefengensis sp. nov. was preparaed by using the ultrasonic methods, the main components in the extracts were separated by using the gradient elution method with RP-HPLC, Yuexu AQ-C18 column (250 mm × 4.6 mm, 5 μm); The positive and negative electro spray ionization (ESI) source was used for determine the chromatographic effluent, the main chromatographic peaks are assigned by Q-TOF. Based on the standards of MS/MS and compared with the reference results, 28 compounds, containing mannitol, adenosine, ergosterol, sitosterol, amino acid, fatty acid, and sugar, were identified. HPLC-ESI-TOF-MS/MS method can qualitatively analyze the main components in O. xuefengensis sp. nov. from the retention time, UV spectrum, precious relative molecular weight, formula, secondary fragment ions, and so on. It is an effective and fast analysis method for determining the active compounds in O. xuefengensis sp. nov.